ObjectiveTo study the contents of CD44 that shared exon variant 5 (CD44v5) in peripheral blood lymphocytes (PBL) of patients with gastric carcinoma and the expression of CD44v5 in tumor tissue and their clinical significance. MethodsThe contents of CD44v5 were determined by FlowCytometry in PBL of 31 patients with gastric carcinoma before surgery and 10 normal controls. Tissue expression of CD44v5 in 33 patients with gastric carcinoma was investigated by immunohistochemistry. ResultsThe contents of CD44v5 were significantly higher in PBL of patients with gastric carcinoma before surgery than those of controls (P＜0.01). Nodepositive gastric cancer patients showed significantly elevated contents of CD44v5 in PBL in comparison with nodenegative gastric cancer (P＜0.01）. Significant correlations were noted between the contents of CD44v5 in PBL of patients with gastric carcinoma before surgery and tumor size, depth of invasion, lymph node metastasis and different the Vnion International Centre Le Cancer (VICC) stages of tumor (P＜0.05). The expression of CD44v5 could be detected in 69.7% of tumor tissue,but was not detected in adjacent normal gastric mucosa. Significant correlations were noted between CD44v5 expression and depth of invasion,and lymph node metastasis.The presence of CD44v5 protein was correlated with the lymph node involvement rate. Conclusion CD44v5 in PBL or tumor tissue may be useful as a metastatic marker. It may be of important clinical value in the diagnosis of metastasis and judgement of development for the patients with gastric cancer.
ObjectiveTo determine the nuclear factor kappa B (NFkB) activity in peripheral blood mononuclear cells (PBMC) in patients with acute cholangitis of severe type (ACST) and correlate the degree of NFkB activation with severity of biliary tract infection and clinical outcome.MethodsTwenty patients with ACST were divided into survivor group (14 cases) and nonsurvivor group (6 cases). Other 10 patients undergoing elective gastrectomy or inguinal hernia repair were selected as control group. Peripheral blood samples were taken 24 hours after operation, PBMC was separated and nuclear proteins were isolated from PBMC, and NFkB was determined with electrophoretic mobility shift assay (EMSA). The levels of TNFα, IL6 and IL10 in plasma were determined by using an enzymelinked immunoassay (ELISA). ResultsThe NFkB activity was 5.02±1.03, 2.98±0.51 and 1.02±0.34 respectively in three groups. It was increased in all patients with ACST, versus the control group (P＜0.05), and the patients of nonsurvivor group had higher levels of NFkB activation than those of survivor group (P＜0.05). The levels of TNFα and IL6 were (496.28±52.35) ng/L and (578.13±67.72) ng/L in nonsurvivor group; (284.47±39.41) ng/L and (318.67±34.92) ng/L in survivor group; (89.43±10.39) ng/L and (101.27±13.47) ng/L in control group. All patients with ACST had increased levels of TNFα and IL6, which were many fold greater than that of control group, and there was an evidence of significantly higher levels in nonsurvivor group than in survivor group (P＜0.05). All patients had also increased levels of IL10 as compared to control group (P＜0.05), but the IL10 concentrations in plasma were not significantly higher in nonsurvivors than that of in those survivors (Pgt;0.05). ConclusionNFkB activation in PBMCs in patients with ACST
ObjectiveTo investigate the clinical significancy of K-ras gene mutation in peripheral blood free DNA in patients with non-small cell lung cancer (NSCLC). MethodsA total of 242 patients pathologically diagnosed with NSCLC in the Third People's Hospital of Chengdu were recruited between January 2013 and August 2015. Both tumor tissues and peripheral blood free DNA were collected for detection of K-ras gene mutation by mutant-enriched liquidchip technology. The detection rate was compared between these two kinds of samples. ResultsIn tumor tissues, the K-ras gene mutation was detected in 12 cases with a positive rate of 4.96%. While in peripheral blood samples, the K-ras gene mutation was detected in 10 cases with a positive rate of 4.13%. The detection yield of K-ras gene mutation in peripheral blood had a good consistency with that of lung cancer tissues (Kappa value=0.81). ConclusionK-ras in peripheral blood plasma free DNA can be a surrogate marker for tumor tissues' K-ras gene mutation in screening patients with NSCLC.
Objective To detect expression of CK20 mRNA in peripheral blood of patients with colorectal carcinoma and its clinical significance. Methods Using the reverse transcriptase-polymerase chain reaction (RT-PCR),CK20 mRNA expression was examined in peripheral blood from 42 patients with colorectal carcinoma before and after operation, 20 healthy volunteers, 20 fresh colorectal carcinoma samples. Results The positive expression rates of CK20 mRNA were 45.24%(19/42) and 33.33%(14/42) before and after operation in 42 colorectal carcinoma patients respectively. All 20 fresh colorectal carcinoma samples revealed expression of CK20 mRNA, but the 20 normal blood samples did not. Conclusion The detection of CK20 mRNA in peripheral blood is helpful to early diagnose, assess the prognosis and make a correct treatment of colorectal carcinoma.
Objective To determine the application values of gene chip technique in cardiovascular surgical clinical and research work. Microarray for gene expression profiles was used to screen out the differentially expressed genes during cardiopulmonary bypass(CPB) in peripheral blood mononuclear cell. By doing these, it was hoped that some clues in inflammatory response during CPB could be found out. Methods The patients’ oxygenated bloods were drawn immediately before onset and termination of CPB. Peripheral blood mononuclear cell (PBMC) were obtained from heparinised blood by Ficoll gradient centrifugation. The differentially expression was measured using BD AtlasTM cDNA Expression Arrays. The candidate genes were corroborated by semiquantitative reverse transcriptionpolymerase chain reaction (RT-PCR). Results Gene chip technique was successfully used in CPB study. The gene expression profiles of cytokines of PBMC during CPB were screened out. Interleukin 6 and Wnt5a were the differentially expressed genes. But the validity using semiquantitative RT-PCR found no statistically difference(P=0.888,0.135). Conclusion Microarray technique has positive application values in the study of cytokines during CPB. cDNA microarray for gene expression profiles can primarily screen out differentially expression genes during CPB. These genes may be engaged in inflammation and other pathophysiological reactions during CPB. PBMC is not the major source of cytokines during CPB.
ObjectTo investigate the pathogenesis of drug-resistant epilepsy by examining the expression of mRNA and protein of Cell Division Cycle 42 GTP-binding protein (Cdc42), Neural Wiskott-Aldrich Syndrome Protein (N-WASP) and Actin-related protein 2/3(Arp2/3) in peripheral blood of patients with drug-resistant epilepsy (DRE).MethodsSeventy two essential epilepsy patients who were attended at outpatients and inpatients in the Department of Neurology of the Affiliated Hospital of Youjiang Medical University for Nationalities were selected from October 2016 to October 2018. According to the 2010 International League Against Epilepsy’s definition of Drug-Resistant Epilepsy, the patients were divided into 2 groups: 32 patients with DRE were defined as DRE group, 40 patients with anti-epilepsy drugs (AEDs) well controlled were defined as the well controlled group. Thirty two healthy persons were selected as control group. The expression of mRNA and protein of Cdc42, N-WASP and Arp2/3 in peripheral blood were measured by quantitative real-time PCR (RT-qPCR) and Western blot(WB). Experimental data were analyzed by ANOVA or rank-sum test.ResultsCompared with well-controlled group and healthy persons group, Cdc42, N-WASP, Arp2/3 in DRE group were significantly increased, the differences were statistically significant (P<0.05). Compared with the control group, Cdc42, N-WASP, Arp2/3 in well-controlled group were significantly increased, with statistically significant differences (P<0.05).ConclusionThe expression of Cdc42, N-WASP, Arp2/3 in peripheral blood of patients with DRE significantly increased, being closely related to the occurrence and development of DRE, and used as indicators in peripheral blood predicting the occurrence of DRE.
Effect of radical operation on expression of interleukin-2(IL-2)mRNA and production of IL-2 were markedly reduced preoperatively and four weeks after operation,expression of IL-2 mRNA significantly enhanced,but it was still lower than that in the normal group.Production of IL-2 nearly reached normal level,When PBL was activated by phytohemagglutinin(PHA),expresseion of IL-2 mRNA and production of IL-2 were much higher than that in non-activated PBL.These results suggested that expression of IL-2 were much higher than that in non-activated PBL.These results suggested that expression of IL-2 mRNA and production of IL-2 are dificient in gastric cancer patients,and radical surgery will help them to recover and they can also be improved through activation with PHA.
ObjectiveTo research the effect of recombinant adenovirus-bone morphogenetic protein 12 (Ad-BMP-12) transfection on the differentiation of peripheral blood mesenchymal stem cells (MSCs) into tendon/ligament cells. MethodsPeripheral blood MSCs were isolated from New Zealand rabbits (3-4 months old) and cultured in vitro until passage 3. The recombinant adenoviral vector system was prepared using AdEasy system, then transfected into MSCs at passage 3 (transfected group); untransfected MSCs served as control (untransfected group). The morphological characteristics and growth of transfected cells were observed under inverted phase contrast microscope. The transfection efficiency and green fluorescent protein (GFP) expression were detected by flow cytometry (FCM) and fluorescence microscopy. After cultured for 14 days in vitro, the expressions of tendon/ligament-specific markers were determined by immunohistochemistry and real-time fluorescent quantitative PCR. ResultsGFP expression could be observed in peripheral blood MSCs at 8 hours after transfection. At 24 hours after transfection, the cells had clear morphology and grew slowly under inverted phase contrast microscope and almost all expressed GFP at the same field under fluorescence microscopy. FCM analysis showed that the transfection efficiency of the transfected group was 99.57%, while it was 2.46% in the untransfected group. The immunohistochemistry showed that the expression of collagen type Ι gradually increased with culture time in vitro. Real-time fluorescent quantitative PCR results showed that the mRNA expressions of the tendon/ligament-specific genes (Tenomodulin, Tenascin-C, and Decorin) in the transfected group were significantly higher than those in untransfected group (0.061±0.013 vs. 0.004±0.002, t=-7.700, P=0.031; 0.029±0.008 vs. 0.003±0.001, t=-5.741, P=0.020; 0.679±0.067 vs. 0.142±0.024, t=-12.998, P=0.000). ConclusionAd-BMP-12 can significantly promote differentiation of peripheral blood MSCs into tendon/ligament fibroblasts and enhance the expressions of tendon/ligament-specific phenotypic differentiation, which would provide the evidence for peripheral blood MSCs applied for tendon/ligament regeneration.
From March 1979 to February 1987, 500 cases of firearm wounds of blood vessels were treated. Of them, 465 cases were recovered, 15 cases were disabled, 13 cases had amputation, and 7 cases died. The article presented the clinic materials. The following problems were discussed: (1) The characteristics of firearm wounds of blood vessels. (2) Emergency treatment of injuries of major blood vessels of limbs. (3) Indications of repair of blood vessels. (4) The methods of repair of defect in blood vessel. (5)Factors influencing the survival of extremities, and (6) Active prevention and treatment of complication.
Objective To study the expression of NLRP3 inflammasome and its downstream inflammatory factors in patients with chronic obstructive pulmonary disease (COPD) and healthy controls, and to reveal the effect and significance of NLRP3 inflammasome in the pathogenesis of COPD. Methods Forty patients with acute exacerbation COPD (AECOPD) who were hospitalized from November 2016 to May 2017 were recruited in the AECOPD group, and recruited in the stable COPD group when they entered the stable stage. Forty healthy individuals were recruited in the control group. General information and peripheral blood were collected from each subject. The levels of NLRP3 mRNA and caspase-1 mRNA in peripheral blood mononuclear cells were measured by real-time PCR. The levels of IL-18 and IL-1β were measured by enzyme-linked immunosorbent assay. Results The levels of NLRP3 mRNA, IL-18 and IL-1β in the AECOPD patients were significantly higher than those in the stable COPD group [2.11±0.77, 12.79 (7.10, 43.13) pg/ml, 17.02 (8.36, 52.21) pg/ml vs. 1.60±0.44, 10.66 (6.32, 18.59) pg/ml, 13.34 (7.07, 16.89) pg/ml, all P<0.05] . The levels of NLRP3 mRNA, IL-18 and IL-1β in the AECOPD patients were significantly higher than those in the control group [2.11±0.77, 12.79 (7.10, 43.13) pg/ml, 17.02 (8.36, 52.21) pg/mlvs. 1.00±0.49, 6.29 (4.73, 7.93) pg/ml, 5.93 (4.81, 9.67) pg/ml, all P<0.05]. The levels of NLRP3 mRNA, IL-18 and IL-1β were significantly higher in the stable COPD group than the control group [1.60±0.44, 10.66 (6.32, 18.59) pg/ml, 13.34 (7.07, 16.89) pg/mlvs. (1.00±0.49, 6.29 (4.73, 7.93) pg/ml, 5.93 (4.81, 9.67) pg/ml, all P<0.05]. Correlation analysis showed that the plasma IL-18 level was positive correlated with leukocyte count and neutrophil percentage in the AECOPD group (r=0.372, P<0.05;r=0.386, P<0.05). The expression of NLRP3 mRNA in the AECOPD group and stable COPD group were positively correlated with the CAT score (r=0.387, P<0.05;r=0.399, P<0.05) . Conclusion NLRP3 inflammasome is involved in the inflammatory response in COPD patients.